Chloramine-T Margin-of-Safety Estimates for Fry, Fingerling, and Juvenile Rainbow Trout

Abstract

Chloramine-T (CLT) is a candidate for approval for use in U.S. aquaculture to control mortality in freshwater-reared salmonids caused by bacterial gill disease (causative agent, Flavobacterium branchiophilum). The proposed treatment regimen is to administer CLT at 12–20 mg/L in a static or flow-through bath for 60 min/d on three alternate or consecutive days. To estimate a CLT margin of safety, defined as the highest dosing regimen above the proposed maximum therapeutic regimen at which no adverse effects are observed, we conducted seven experiments with fry, fingerling, and juvenile rainbow trout Oncorhynchus mykiss that examined mortality and an eighth experiment that examined mortality, gross pathology, and histopathology after CLT exposure. In each experiment, triplicate groups of fish were exposed to a range of CLT concentrations representing 0, 1, 1.5, 2, 2.5, 3, 3.5, 4, or 5× the highest proposed dose (20 mg/L) for 3× the proposed treatment duration (60 min) on three alternate or consecutive days at 8°C or 14°C. The survival of fry and fingerlings was unaffected by exposure to CLT concentrations as high as 100 and 60 mg/L, respectively (survival = 97.3–100%). Although the survival of juvenile fish was unaffected by exposure to 20 mg/L, exposure to higher CLT concentrations significantly reduced survival (≤10.0% at 100 mg/L). Across experiments, 92% of all mortalities occurred within 20 h of the first exposure to CLT. The histopathological changes of most concern were associated with gill tissues, but these were evident only in moribund fish exposed to doses of 60 mg/L or higher. Based on analysis of the survival data, the margin-of-safety estimates were approximately 100 mg/L for rainbow trout fry, at least 60 mg/L for fingerlings, and 50–60 mg/L for juveniles. Tissue responses to CLT at these concentrations were minor and did not warrant decreasing these estimates.

ACKNOWLEDGMENTS

U.S. Fish and Wildlife Service biologists Bonnie Johnson, Andy Gray, and Molly Bowman (Aquatic Animal Drug Approval Partnership Program), and Chris Vialponda, Greg Kindschi, and Ron Zitzow (Bozeman Fish Technology Center), served as masked or nonmasked study participants. U.S. Fish and Wildlife Service biologists Crystal Hudson and Tammy Weiss (Bozeman Fish Health Center, Bozeman, Montana) performed bacterial and viral analyses for fish health evaluations. Mark Gaikowski, U.S. Geological Survey, Upper Midwest Environmental Sciences Center, La Crosse, Wisconsin, assisted with statistical analysis of survival data. John Borkowski, Department of Mathematical Sciences, Montana State University, calculated standard deviations for chloramine-T doses administered. Jesse Trushenski, Southern Illinois University, and Beth MacConnell, USFWS retired, provided critical review of this manuscript.